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Vol. 11, Issue 5, 1765-1774, May 2000
Department of Physiology, University of Massachusetts Medical
School, Worcester, Massachusetts 01655
In animal cells, positioning of the mitotic spindle is crucial for
defining the plane of cytokinesis and the size ratio of daughter cells.
We have characterized this phenomenon in a rat epithelial cell line
using microscopy, micromanipulation, and microinjection. Unmanipulated
cells position the mitotic spindle near their geometric center, with
the spindle axis lying roughly parallel to the long axis of the cell.
Spindles that were initially misoriented underwent directed rotation
and caused a delay in anaphase onset. To gain further insight into this
process, we gently deformed cells with a blunted glass needle to change
the spatial relationship between the cortex and spindle. This
manipulation induced spindle movement or rotation in metaphase and/or
anaphase, until the spindle reached a proper position relative to the
deformed shape. Spindle positioning was inhibited by either treatment
with low doses of nocodazole or microinjection of antibodies against dynein, apparently due to the disruption of the organization of dynein
and/or astral microtubules. Our results suggest that mitotic cells
continuously monitor and maintain the position of the spindle relative
to the cortex. This process is likely driven by interactions among
astral microtubules, the motor protein dynein, and the cell cortex and
may constitute part of a mitotic checkpoint mechanism.
Online version of this article contains video
material for Figures 1, 3, 4, 7, and 9. Online version available at
www.molbiolcell.org.
*
Corresponding author. E-mail address:
yuli.wang{at}umassmed.edu.
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